限氧与通氧条件下米根霉.pdfVIP

中国农业科学 2006,39(6):1228-1232 Scientia Agricultura Sinica 限氧与通氧条件下米根霉ADH突变株的碳代谢特性 郑 志，姜绍通，罗水忠，李兴江，潘丽军，赵 辛 （合肥工业大学生物与食品工程学院，合肥 230009 ） 摘要：【目的】乙醇脱氢酶（ADH）是米根霉发酵生产L-乳酸代谢途径中的支路关键酶，选育米根霉ADH突变 株，降低ADH 活力，可以提高乳酸转化率。【方法】利用亚硝基胍（NTG）诱变米根霉As3.3462，在含丙烯醇的YPD 选择培养基上筛选获得ADH 突变株，研究了突变菌株在限氧与通氧条件下的乙醇、乳酸、ADH与 L-乳酸脱氢酶（LDH） 的特性。【结果】 获得11株ADH突变株，其中mut-2突变株在通氧和限氧条件下对乳酸的转化率比出发菌株分别 提高了84.6%和175%，而对乙醇的转化分别下降了39.5%和59.8%。mut-2突变株的最大ADH活力在通氧和限氧条 件下均比出发菌株降低，而LDH活力在限氧条件下显著增加，通氧条件下变化不明显。【结论】氧气的供给对米根 霉发酵产 L-乳酸的产物组成比例影响很大，限氧发酵条件下ADH活力提高，乙醇积累。结果还表明，通过筛选米 根霉ADH突变株，是获得 L-乳酸高产菌株的一种简单快速的有效手段。 关键词：米根霉；ADH；乙醇；L-乳酸 Carbon Metabolism Characteristics of Rhizopus oryzae ADH Mutant under Aeration and Oxygen-limiting Fermentations ZHENG Zhi, JIANG Shao-tong, LUO Shui-zhong, LI Xing-jiang, PAN Li-jun, ZHAO Xin (School of Biotechnology and Food Engineering, Hefei University of Technology, Hefei 230009) Abstract: 【Objective 】Ethanol dehydrogenase (ADH) is an important branch pathway enzyme that catalyzes the transformation of ethanol from pyruvate in Rhizopus oryzae, which decrease the conversion rate of glucose to L-lactic acid. So, screening the mutants with lower ADH activity may markedly increase lactate production. 【Method 】 Mutation of Rhizopus oryzae As3.3462 was induced with N-methyl-N-nitro-N-nitrosoguanidine (NTG) and mutants with lower ADH activity were screened out, then the characteristics of mutant under aeration and oxygen-limiting fermentations were investigated. 【Result 】 There were 11 Rhizopus oryzae strains of ADH mutants were obtained. After preliminary fermentation, mut-2 was selected for further study on the fermentation characteristics because of its high lactate conversion